Dye coupling among neurons has been shown to reflect electrotonic coupling. Recent work in retina has revealed that the incidence of coupling is greater when estimated by neurobiotin (NB) transfer than by Lucifer yellow (LY). Several previous studies have shown that the incidence of LY coupling among supraoptic nucleus (SON) neurons of lactating rats is 2- to 4-fold higher than is observed in virgin females. We compared the incidence of coupling among SON neurons following simultaneous injections of LY and NB into the same cells in slices from virgin or lactating rats. As seen in previous studies, there were 4-fold more LY-coupled neurons per injection in lactating as compared to virgin rats. Under both conditions, the numbers of NB-coupled neurons per injection were 4-fold greater than was observed for LY; possible mechanisms are discussed. Individual NB-filled neurons were coupled to as many as 10 other cells distributed over a large area of the SON. These results confirm previous findings of more coupling in lactating than virgin SONs, and suggest that both the incidence and spatial extent of interneuronal coupling are greater and thus probably more important functionally than has been heretofore appreciated.